• Title/Summary/Keyword: Small molecules

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The Ability of Anti-tumor Necrosis Factor Alpha(TNF-${\alpha}$) Antibodies Produced in Sheep Colostrums

  • Yun, Sung-Seob
    • 한국유가공학회:학술대회논문집
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    • 2007.09a
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    • pp.49-58
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    • 2007
  • Inflammatory process leads to the well-known mucosal damage and therefore a further disturbance of the epithelial barrier function, resulting abnormal intestinal wall function, even further accelerating the inflammatory process[1]. Despite of the records, etiology and pathogenesis of IBD remain rather unclear. There are many studies over the past couple of years have led to great advanced in understanding the inflammatory bowel disease(IBD) and their underlying pathophysiologic mechanisms. From the current understanding, it is likely that chronic inflammation in IBD is due to aggressive cellular immune responses including increased serum concentrations of different cytokines. Therefore, targeted molecules can be specifically eliminated in their expression directly on the transcriptional level. Interesting therapeutic trials are expected against adhesion molecules and pro-inflammatory cytokines such as TNF-${\alpha}$. The future development of immune therapies in IBD therefore holds great promises for better treatment modalities of IBD but will also open important new insights into a further understanding of inflammation pathophysiology. Treatment of cytokine inhibitors such as Immunex(Enbrel) and J&J/Centocor(Remicade) which are mouse-derived monoclonal antibodies have been shown in several studies to modulate the symptoms of patients, however, theses TNF inhibitors also have an adverse effect immune-related problems and also are costly and must be administered by injection. Because of the eventual development of unwanted side effects, these two products are used in only a select patient population. The present study was performed to elucidate the ability of TNF-${\alpha}$ antibodies produced in sheep colostrums to neutralize TNF-${\alpha}$ action in a cell-based bioassay and in a small animal model of intestinal inflammation. In vitro study, inhibitory effect of anti-TNF-${\alpha}$ antibody from the sheep was determined by cell bioassay. The antibody from the sheep at 1 in 10,000 dilution was able to completely inhibit TNF-${\alpha}$ activity in the cell bioassay. The antibodies from the same sheep, but different milkings, exhibited some variability in inhibition of TNF-${\alpha}$ activity, but were all greater than the control sample. In vivo study, the degree of inflammation was severe to experiment, despite of the initial pilot trial, main trial 1 was unable to figure out of any effect of antibody to reduce the impact of PAF and LPS. Main rat trial 2 resulted no significant symptoms like characteristic acute diarrhea and weight loss of colitis. This study suggested that colostrums from sheep immunized against TNF-${\alpha}$ significantly inhibited TNF-${\alpha}$ bioactivity in the cell based assay. And the higher than anticipated variability in the two animal models precluded assessment of the ability of antibody to prevent TNF-${\alpha}$ induced intestinal damage in the intact animal. Further study will require to find out an alternative animal model, which is more acceptable to test anti-TNF-${\alpha}$ IgA therapy for reducing the impact of inflammation on gut dysfunction. And subsequent pre-clinical and clinical testing also need generation of more antibody as current supplies are low.

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Anti-inflammatory Activity of Antimicrobial Peptide Zophobacin 1 Derived from the Zophobas atratus (아메리카왕거저리 유래 항균 펩타이드 조포바신 1의 항염증활성)

  • Shin, Yong Pyo;Lee, Joon Ha;Kim, In-Woo;Seo, Minchul;Kim, Mi-Ae;Lee, Hwa Jeong;Baek, Minhee;Kim, Seong Hyun;Hwang, Jae Sam
    • Journal of Life Science
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    • v.30 no.9
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    • pp.804-812
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    • 2020
  • The giant mealworm beetle, Zophobas atratus (Coleoptera: Tenebrionidae) has been used as a protein source for small pets and mammals. Recently, it was temporarily registered in the list of the Food Code. We previously performed an in silico analysis of the Zophobas atratus transcriptome to identify putative antimicrobial peptides and identified several antimicrobial peptide candidates. Among them, we assessed the antimicrobial and anti-inflammatory activities of zophobacin 1 that was selected bio-informatically based on its physicochemical properties against microorganisms and mouse macrophage Raw264.7 cells. Zophobacin 1 showed antimicrobial activities against microorganisms without inducing hemolysis and decreased the nitric oxide production of the lipopolysaccharide-induced Raw264.7 cells. Moreover, ELISA and Western blot analysis revealed that zophobacin 1 reduced expression levels of pro-inflammatory enzymes such as inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). We also investigated expression of pro-inflammatory cytokines (interleukin-6 and interleukin-1β) production through quantitative real time-PCR and ELISA. Zophobacin 1 markedly reduced the expression level of cytokines through the regulation of mitogen-activated protein kinases (MAPKs) and nuclear factor kappa B (NF-κB) signaling. We confirmed that zophobacin 1 bound to bacterial cell membranes via a specific interaction with lipopolysaccharides. These data suggest that zophobacin 1 could be promising molecules for development as antimicrobial and anti-inflammatory therapeutic agents.

INTRINSIC NMR ISOTOPE SHIFTS OF CYCLOOCTANONE AT LOW TEMPERATURE (저온에서의 싸이클로옥타논에 대한 고유동위원소 효과)

  • Jung, Miewon
    • Analytical Science and Technology
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    • v.7 no.2
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    • pp.213-224
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    • 1994
  • Several isotopomers of cyclooctanone were prepared by selective deuterium substitution. Intrinsic isotope effects on $^{13}C$ NMR chemical shifts of these isotopomers were investigated systematically at low temperature. These istope effects were discussed in relation to the preferred boat-chair conformation of cyclooctanone. Deuterium isotope effects on NMR chemical shifts have been known for a long time. Especially in a conformationally mobile molecule, isotope perturbation could affect NMR signals through a combination of isotope effects on equilibria and intrinsic effects. The distinction between intrinsic and nonintrinsic effects is quite difficult at ambient temperature due to involvement of both equilibrium and intrinsic isotope effects. However if equilibria between possible conformers of cyclooctanone are slowed down enough on the NMR time scale by lowering temperature, it should be possible to measure intrinsic isotope shifts from the separated signals at low temperature. $^{13}C$ NMR has been successfully utilized in the study on molecular conformation in solution when one deals with stable conformers or molecules were rapid interconversion occurs at ambient temperature. The study of dynamic processes in general requires analysis of spectra at several temperature. Anet et al. did $^1H$ NMR study of cyclooctanone at low temperature to freeze out a stable conformation, but were not able initially to deduce which conformation was stable because of the complexity of alkyl region in the $^1H$ NMR spectrum. They also reported the $^1H$ and $^{13}C$ NMR spectra of the $C_9-C_{16}$ cycloalkanones with changing temperature from $-80^{\circ}C$ to $-170^{\circ}C$, but they did not report a variable temperature $^{13}C$ NMR study of cyclooctanone. For the analysis of the intrinsic isotope effect with relation to cylooctanone conformation, $^{13}C$ NMR spectra are obtained in the present work at low temperatures (up to $-150^{\circ}C$) in order to find the chemical shifts at the temperature at which the dynamic process can be "frozen-out" on the NMR time scale and cyclooctanone can be observed as a stable conformation. Both the ring inversion and pseudorotational processes must be "frozen-out" in order to see separate resonances for all eight carbons in cyclooctanone. In contrast to $^1H$ spectra, slowing down just the ring inversion process has no apparent effects on the $^{13}C$ spectra because exchange of environments within the pairs of methylene carbons can still occur by the pseudorotational process. Several isotopomers of cyclooctanone were prepared by selective deuterium substitution (fig. 1) : complete deuterium labeling at C-2 and C-8 positions gave cyclooctanone-2, 2, 8, $8-D_4$ : complete labeling at C-2 and C-7 positions afforded the 2, 2, 7, $7-D_4$ isotopomer : di-deuteration at C-3 gave the 3, $3-D_2$ isotopomer : mono-deuteration provided cyclooctanone-2-D, 4-D and 5-D isotopomers : and partial deuteration on the C-2 and C-8 position, with a chiral and difunctional case catalyst, gave the trans-2, $8-D_2$ isotopomer. These isotopomer were investigated systematically in relation with cyclooctanone conformation and intrinsic isotope effects on $^{13}C$ NMR chemical shifts at low temperature. The determination of the intrinsic effects could help in the analysis of the more complex effects at higher temperature. For quantitative analysis of intrinsic isotope effects, the $^{13}C$ NMR spectrum has been obtained for a mixture of the labeled and unlabeled compounds because the signal separations are very small.

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Film Dosimetry for Intensity Modulated Radiation Therapy : Dosimetric Evaluation (필름을 사용한 세기변조치료법에 대한 선량측정)

  • Ju Sang Gyu;Yeo Inhwan Jason;Huh Seung Jae;Choi Byung Ki;Park Young Hwan;Ahn Yong Chan;Kim Dae Yong;Kong Young Kun
    • Radiation Oncology Journal
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    • v.20 no.2
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    • pp.172-178
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    • 2002
  • Purpose : X-ray film over responds to low-energy photons in relative photon beam dosimetry because its sensor is based on silver bromide crystals, which are high-Z molecules. This over-response becomes a significant problem in clinical photon beam dosimetry particularly in regions outside the penumbra. In intensity modulated radiation therapy (IMRT), the radiation field is characterized by multiple small fields and their outside-penumbra regions. Therefore, in order to use film dosimetry for IMRT, the nature the source of the over-response in its radiation field need to be known. This study is aimed to verify and possibly improve film dosimetry for IMRT. Materials and Method : Modulated beams were constructed by a combination of five or seven different static radiation fields using 6 MeV X-rays. In order to verify film dosimetry, we used X-ray film and an ion chamber were used to measure the dose profiles at various depths in a phantom. In addition, in order to reduce the over-response, 0.01 inch thick lead filters were placed on both sides of the film. Results : The measured dose profiles showed a film over-response at the outside-penumbra and low dose regions. The error increased with depths and approached 15% at a maximum for the field size of $15{\times}15cm^2$ at 10 cm depth. The use of filters reduced the error to 3%, but caused an under-response of the dose in a perpendicular set-up. Conclusion : This study demonstrated that film dosimetry for IMRT involves sources of error due to its over-response to low-energy Photons. The use of filers can enhance the accuracy in film dosimetry for IMRT. In this regard, the use of optimal filter conditions is recommended.

Impact of Sulfur Dioxide Impurity on Process Design of $CO_2$ Offshore Geological Storage: Evaluation of Physical Property Models and Optimization of Binary Parameter (이산화황 불순물이 이산화탄소 해양 지중저장 공정설계에 미치는 영향 평가: 상태량 모델의 비교 분석 및 이성분 매개변수 최적화)

  • Huh, Cheol;Kang, Seong-Gil;Cho, Mang-Ik
    • Journal of the Korean Society for Marine Environment & Energy
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    • v.13 no.3
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    • pp.187-197
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    • 2010
  • Carbon dioxide Capture and Storage(CCS) is regarded as one of the most promising options to response climate change. CCS is a three-stage process consisting of the capture of carbon dioxide($CO_2$), the transport of $CO_2$ to a storage location, and the long term isolation of $CO_2$ from the atmosphere for the purpose of carbon emission mitigation. Up to now, process design for this $CO_2$ marine geological storage has been carried out mainly on pure $CO_2$. Unfortunately the $CO_2$ mixture captured from the power plants and steel making plants contains many impurities such as $N_2$, $O_2$, Ar, $H_2O$, $SO_2$, $H_2S$. A small amount of impurities can change the thermodynamic properties and then significantly affect the compression, purification, transport and injection processes. In order to design a reliable $CO_2$ marine geological storage system, it is necessary to analyze the impact of these impurities on the whole CCS process at initial design stage. The purpose of the present paper is to compare and analyse the relevant physical property models including BWRS, PR, PRBM, RKS and SRK equations of state, and NRTL-RK model which are crucial numerical process simulation tools. To evaluate the predictive accuracy of the equation of the state for $CO_2-SO_2$ mixture, we compared numerical calculation results with reference experimental data. In addition, optimum binary parameter to consider the interaction of $CO_2$ and $SO_2$ molecules was suggested based on the mean absolute percent error. In conclusion, we suggest the most reliable physical property model with optimized binary parameter in designing the $CO_2-SO_2$ mixture marine geological storage process.

Effect of Nitrogen Impurity on Process Design of $CO_2$ Marine Geological Storage: Evaluation of Equation of State and Optimization of Binary Parameter (질소 불순물이 이산화탄소 해양 지중저장 공정설계에 미치는 영향 평가: 상태방정식의 비교 분석 및 이성분 매개변수 최적화)

  • Huh, Cheol;Kang, Seong-Gil
    • Journal of the Korean Society for Marine Environment & Energy
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    • v.12 no.3
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    • pp.217-226
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    • 2009
  • Marine geological storage of $CO_2$ is regarded as one of the most promising options to response climate change. Marine geological storage of $CO_2$ is to capture $CO_2$ from major point sources, to transport to the storage sites and to store $CO_2$ into the marine geological structure such as deep sea saline aquifer. Up to now, process design for this $CO_2$ marine geological storage has been carried out mainly on pure $CO_2$. Unfortunately the captured $CO_2$ mixture contains many impurities such as $N_2$, $O_2$, Ar, $H_2O$, $SO_x$, $H_2S$. A small amount of impurities can change the thermodynamic properties and then significantly affect the compression, purification and transport processes. In order to design a reliable $CO_2$ marine geological storage system, it is necessary to perform numerical process simulation using thermodynamic equation of state. The purpose of the present paper is to compare and analyse the relevant equations of state including PR, PRBM, RKS and SRK equation of state for $CO_2-N_2$ mixture. To evaluate the predictive accuracy of the equation of the state, we compared numerical calculation results with reference experimental data. In addition, optimum binary parameter to consider the interaction of $CO_2$ and $N_2$ molecules was suggested based on the mean absolute percent error. In conclusion, we suggest the most reliable equation of state and relevant binary parameter in designing the $CO_2-N_2$ mixture marine geological storage process.

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The Origin of Meju Fungi - Fungal Diversity of Soybean, Rice Straw and Air for Meju Fermentation

  • Kim, Dae-Ho;Lee, Jong-kyu;Hong, Seung-Beom
    • 한국균학회소식:학술대회논문집
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    • 2014.10a
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    • pp.32-32
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    • 2014
  • Meju is a brick of dried fermented soybeans and is the core material for Jang such as Doenjang and Ganjang. Jang is produced by addition of salty water to Meju and is considered the essential sauces of authentic Korean cuisine. Meju is fermented by diverse microorganisms such as bacteria, fungi and yeasts. It is known that fungi play an important role in the Meju fermentation and they degrade macromolecules of the soybeans into small nutrient molecules. In previous study, 26 genera and 0 species were reported as Meju fungi. However, it is not comprehensively examined where the fungi present on the Meju are originated. In order to elucidate the origin of the fungi present on the Meju, the mycobiota of 500 samples soybean kernels, 296 rice straw pieces and air samples of Jang factories was determined in 0, 2 and 7 Jang factories respectively. Forty-one genera covering 86 species were isolated from the soybeans and 33 species were identical with the species from Meju. From sodium hypochlorite untreated soybeans, Eurotium herbariorum, Eurotium repens, Cladosporium tenuissimum, Fusarium fujikuroi, Aspergillus oryzae/flavus and Penicillium steckii were the predominant species. In case of sodium hypochlorite-treated soybeans, Eurotium herbariorum, E. repens and Cladosporium tenuissimum were the predominant species. Of the 4 genera and 86 species isolated from soybeans, 3 genera and 33 species were also found in Meju. Thirty-nine genera and 92 species were isolated from the rice straws and 40 species were identical with the species from Meju. Fusarium asiaticum, Cladosporium cladosporioides, Aspergillus tubingensis, A. oryzae, E. repens and Eurotium chevalieri were frequently isolated from the rice straw obtained from many factories. Twelve genera and 40 species of fungi that were isolated in the rice straw in this study, were also isolated from Meju. Especially, A. oryzae, C. cladosporioides, E. chevalieri, E. repens, F. asiaticum and Penicillium polonicum that are abundant species in Meju, were also isolated frequently from rice straw. C. cladosporioides, F. asiaticum and P. polonicum that are abundant in low temperature fermentation process of Meju fermentation, were frequently isolated from rice straw incubated at $5^{\circ}C$ and $25^{\circ}C$, while A. oryzae, E. repens and E. chevalieri that are abundant in high temperature fermentation process of Meju fermentation, were frequently isolated from rice straw incubated at $25^{\circ}C$ and $35^{\circ}C$. This suggests that the mycobiota of rice straw have a large influence in mycobiota of Meju. Thirty-nine genera and 92 species were isolated from the air of Jang factories and 34 species were identical with the species from Meju. In outside air of the fermentation room, Cladosporium sp. and Cladosporium cladosporioides were the dominant species, followed by Cladosporium tenuissimum, Eurotium sp., Phoma sp. Sistotrema brinkmannii, Alternaria sp., Aspergillus fumigatus, Schizophyllum commune, and Penicillium glabrum. In inside air of the fermentation room, Cladosporium sp., Aspergillus oryzae, Penicillium chrysogenum, A. nidulans, Aspergillus sp., C. cladosporioides, Eurotium sp., Penicillium sp., C. tenuissimum, A. niger, E. herbariorum, A. sydowii, and E. repens were collected with high frequency. The concentrations of the genus Aspergillus, Eurotium and Penicillium were significantly higher in inside air than outside air. From this results, the origin of fungi present on Meju was inferred. Of the dominant fungal species present on Meju, Lichtheimia ramosa, Mucor circinelloides, Mucor racemosus, and Scopulariopsis brevicaulis are thought to be originated from outside air, because these species are not or are rarely isolated from rice straw and soybean; however, they were detected outside air of fermentation room and are species commonly found in indoor environments. However, A. oryzae, P. polonicum, E. repens, P. solitum, and E. chevalieri, which are frequently found on Meju, are common in rice straw and could be transferred from rice straw to Meju. The fungi grow and produce abundant spores during Meju fermentation, and after the spores accumulate in the air of fermentation room, they could influence mycobiota of Meju fermentation in the following year. This could explain why concentrations of the genus Aspergillus, Eurotium, and Penicillium are much higher inside than outside of the fermentation rooms.

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Effects of Follicle Cells on the Chymotrypsin Resistance of Mouse Oocytes (난포세포가 생쥐 난자의 Chymotrypsin에 대한 내성에 미치는 영향)

  • Kim, Seong-Im;Bae, In-Ha;Kim, Hae-Kwon;Kim, Sung-Rye
    • Clinical and Experimental Reproductive Medicine
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    • v.26 no.3
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    • pp.407-417
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    • 1999
  • Objective: Mammalian follicle cells are the most important somatic cells which help oocytes grow, mature and ovulate and thus are believed to provide oocytes with various functional and structural components. In the present study we have examined whether cumulus or granulosa cells might playa role in establishing the plasma membrane structure of mouse oocytes during meiotic maturation. Design: In particular the differential resistances of mouse oocytes against chymotrypsin treatment were examined following culture with or without cumulus or granulosa cells, or in these cell-conditioned media. Results: When mouse denuded oocytes, freed from their surrounding cumulus cells, were cultured in vitro for $17{\sim}18hr$ and then treated with 1% chymotrypsin, half of the oocytes underwent degeneration within 37.5 min ($t_{50}=37.5{\pm}7.5min$) after the treatment. In contrast cumulus-enclosed oocytes showed $t_{50}=207.0$. Similarly, when oocytes were co-cultured with cumulus cells which were not associated with the oocytes but present in the same medium, the $t_{50}$ of co-cultured oocytes was $177.5{\pm}13.1min$. Furthermore, when oocytes were cultured in the cumulus cell-conditioned medium, $t_{50}$ of these oocytes was $190.0{\pm}10.8min$ whereas $t_{50}$ of the oocytes cultured in M16 alone was $25.5{\pm}2.9min$. Granulosa cell-conditioned medium also increased the resistance of oocytes against chymotrypsin treatment such that $t_{50}$ of oocytes cultured in granulosa cell-conditioned medium was $152.5{\pm}19.0min$ while that of oocytes cultured in M16 alone was $70.0{\pm}8.2min$. To see what molecular components of follicle cell-conditioned medium are involved in the above effects, the granulosa cell-conditioned medium was separated into two fractions by using Microcon-10 membrane filter having a 10 kDa cut-off range. When denuded oocytes were cultured in medium containing the retentate, $t_{50}$ of the oocytes was $70.0{\pm}10.5min$. In contrast, $t_{50}$ of the denuded oocytes cultured in medium containing the filtrate was $142.0{\pm}26.5min$. $T_{50}$ of denuded oocytes cultured in medium containing both retentate and filtrate was $188.0{\pm}13.6min$. However, $t_{50}$ of denuded oocytes cultured in M16 alone was $70.0{\pm}11.0min$ and that of oocytes cultured in whole granulosa cell-conditioned medium was $156.0{\pm}27.9min$. When surface membrane proteins of oocytes were electrophoretically analyzed, no difference was found between the protein profiles of oocytes cultured in M16 alone and of those cultured in the filtrate. Conclusions: Based upon these results, it is concluded that mouse follicle cells secrete a factor(s) which enhance the resistance of mouse oocytes against a proteolytic enzyme treatment. The factor appears to be a small molecules having a molecular weight less than 10 kDa.

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Role of HOXA Gene in Human Endometrial Decidualization (인간 자궁내막의 탈락막화에서 HOXA10 유전자의 역할)

  • Lee, Chang-Se;Park, Dong-Wook;Park, Chan-Woo;Kim, Tae-Jin
    • Clinical and Experimental Reproductive Medicine
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    • v.37 no.3
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    • pp.207-216
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    • 2010
  • Objective: This study was performed to clarify the role of HomeoboxA (HOXA) and its related signaling molecules in the decidualization of primary cultured endometrial cells. Methods: Human endometrial tissues were obtained by curettage of hysterectomy specimens from patients with conditions other than endometrial diseases. Tissues were minced and digested with Trypsin-EDTA for 20 min, $37^{\circ}C$. Cells were cultured with DMEM/F12 medium in $37^{\circ}C$, 5% $CO_2$ incubator for 24 hrs. Cells were treated with HOXA10 siRNA and added transforming growth factor (TGF)-${\beta}1$ (10 ng/mL) for 48 hrs to induces decidualization in vitro. Reverse transcription polymerase chain reaction analysis was accomplished to observe the expression of HOXA10, prolactin, cyclooxygenase (COX)-2, peroxisome proliferatoractivated receptor (PPAR)-$\gamma$, and wingless-type MMTV integration site family (Wnt). Results: HOXA10 expression was increased (1.8 fold vs. non-treated control) in TGF-${\beta}1$ treated cells. Decidualization marker, prolactin, was significantly increased in TGF-${\beta}1$ treated cells compared with HOXA10 siRNA treated cells. Endometrial cell differentiation marker, COX-2 was down-regulated by HOXA10 siRNA even if cells were treated with TGF-${\beta}1$. Wnt4 was down-regulated by treated with HOXA10 siRNA, this expression patters was not changed by TGF-${\beta}1$. Expression of PPAR-$\gamma$ was down regulated by TGF-${\beta}1$ in regardless of HOXA10 siRNA treatment. Conclusion: TGF-${\beta}1$ which is induced by progesterone in endometrial epithelial cells may induces stromal cell decidualization via HOXA10 and Wnt signaling cascade.

Promotion effects of steam-dried Betula platyphylla extract on hair regrowth (자작나무 증포 추출물의 발모 촉진 효과)

  • Ahn, Jeong Won;Jang, Su Kil;Jo, Bo Ram;Kim, Hyun Soo;Jeoung, Eui Young;Hillary, Kithenya;Yoo, Yeong Min;Joo, Seong Soo
    • Korean Journal of Food Science and Technology
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    • v.54 no.1
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    • pp.43-51
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    • 2022
  • Regulation of the hair follicle cycle in association with dermal papilla cells is one of the most interesting targets for promoting hair regrowth. In this study, we examined whether steam-dried Betula platyphylla extracts (BPE) promote hair growth by upregulating in vitro and in vivo responses of dermal papilla cells. The data showed that BPE3 contained high amounts of phenolic compounds with higher antioxidant effects and increased hair growth-related genes, including fibroblast growth factor7 and Wnt7b, in dermal papilla cells. Notably, BPE3 effectively enhanced the formation of hair follicles by increasing FGF7, Wnt7b, and vascular endothelial growth factor in C57BL/6N dorsal skins. Additionally, BPE3 significantly decreased the expression of inflammatory repertoires, inducible nitric oxide synthase, interleukin-6, and cyclooxygenase 2. Several small molecules, such as betulin and unsaturated fatty acids, support the pharmacological activity of BPE3. In conclusion, BPE3 effectively promoted hair growth by activating dermal papilla cells and enhancing hair follicle cycles by attenuating the inflammatory environment in the scalp.